Anti-Human BCL2 (Phospho-Ser70) polyclonal antibody

Donepezil (DNPZ) has shown neuroprotective effect in many disorders. The current study tested the putative retinoprotection provided by donepezil in mouse diabetic retinopathy. Swiss albino mice were allocated to, 1] saline control, 2] diabetic, 3&4] diabetic+DNPZ (1 or 4 mg/kg). After induction of diabetes, mice were maintained for 8 weeks then DNPZ therapy was launched for 28 days. Retinas were isolated and used for histopathology and immunohistochemistry for caspase 3 and the anti-apoptotic protein, B-cell lymphoma 2 (BCl2). Retinas were examined for glutamate, acetylcholine and oxidation markers. Western blot analysis measured inflammatory cytokines, N-methyl-D-aspartate receptors (NMDARs), phosphorylated and total phosphatidylinositol-3 kinase and mTOR, BCl2 and cleaved caspase 3. Significant histopathological changes and decreased thickness were found in diabetic retinas (125.52 +/- 2.85 vs. 157.15 +/- 7.55 in the saline group). In addition, retinal glutamate (2.39-fold), inflammatory cytokines and NMDARs proteins (4.9-fold) were higher in the diabetic retinas. Western blot analysis revealed low ratio of phosphorylated/total PI3K (0.21 +/- 0.043 vs. 1 +/- 0.005) and mTOR (0.18 +/- 0.04 vs. 1 +/- 0.005), low BCl2 (0.28 +/- 0.06 vs. 1 +/- 0.005) and upregulated cleaved caspase 3 (5.18 +/- 1.27 vs. 1 +/- 0.05 in the saline group) versus the saline control. DNPZ ameliorated the histopathologic manifestations and to prevent the decrease in retinal thickness. DNPZ (4 mg/kg) improved phosphorylation of PI3K (0.76 +/- 0.12 vs. 0.21 +/- 0.04) and mTOR (0.59 +/- 0.09 vs. 0.18 +/- 0.04) and increased BCl2 (0.75 +/- 0.08 vs. 0.28 +/- 0.06) versus the diabetic control group. This study explained the retinoprotective effect of DNPZ in mouse diabetic retinopathy and highlighted that mitigation of excitotoxicity, improving phosphorylation of PI3K/mTOR and increasing BCl2 contribute to this effect.

Astaxanthin is a member of the carotenoid family well known for its anti-cancer, anti-diabetic, anti-inflammatory and antioxidant nature. This study was designed to investigate the effects of astaxanthin supplementation of the extender (buffer 2) on post-thaw dog semen quality. Semen from four healthy dogs was collected by digital manipulation twice a week. The ejaculates were pooled, washed, divided into four equal aliquots, diluted with the extender supplemented with different concentrations of astaxanthin (0, 0.5, 1 and 2 mu M) and cryopreserved. The results showed that 1 mu M astaxanthin was the optimum concentration that led to significantly higher (p < .05) post-thaw motility, kinematic parameters and viability than the other groups. In comparison with the control group, sperm samples supplemented with 1 mu M astaxanthin showed significantly higher (p < .05) sperm counts with intact membranes (55.7 +/- 0.6% vs. 51.3 +/- 0.9%), intact acrosome (58.4 +/- 0.7% vs. 53.5 +/- 0.6%), active mitochondria (54.9 +/- 0.5% vs. 42.6 +/- 0.6%) and normal chromatin (67.6 +/- 0.9% vs. 61.7 +/- 0.6%). Furthermore, astaxanthin-supplemented samples showed significantly lower expression levels (p < .05) of pro-apoptotic (BAX), oxidative induced DNA damage repair (OGG1), oxidative stress-related (ROMO1) genes and higher expression levels of anti-apoptotic (BCL2), and sperm acrosome-associated (SPACA3) genes compared to the control. Thus, supplementation of 1 mu M astaxanthin in semen extender results in improved freeze-thaw sperm quality of the dog.